Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Construction of Bioluminescent Escherichia coli from lux Operon and Heat Shock Promoter for the Detection of Toxic Substances

lux Operon과 Heat Shock Promoter 유전자 재조합을 통한 독성물질 탐지용 대장균의 개발

  • 유승오 (경희대학교 생명과학부 식품가공학과) ;
  • 이은관 (경희대학교 생명과학부 식품가공학과) ;
  • 김현숙 (경희대학교 생명과학부 식품가공학과) ;
  • 정계훈 (경희대학교 생명과학부 식품가공학과) ;
  • 전억한 (경희대학교 생명과학부 식품가공학과)
  • Published : 1999.08.01
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Abstract

In order to use heat shock promoter for the detection of toxic substances, dnaK promoter was amplified from E. coli genomic DNA by using a polymerase chain reaction(PCR) followed by sequencing and sub-cloning into the multi-cloning site of the plasmid, pUCD615. The pUCD615 is a broad-host-range vector containing promoterless lux operon originated from V.fischeri. The recombinant plasmid was transfered to E. coli DH5$\alpha$ through electroporation. The recombinant E. coli showed several patterns of bioluminescent responses to ethanol stress. The bioluminescent E. coli also showed responses to other toxic substances including FeK3(CN)6, CdCl2, p-nitrophenol and HgCl2. The increases of RLU(Relative Light Unit) were observed at 100ppm of FeK3(CN)6, 10ppm and 100ppm and 100ppm of CdCl2, 1ppm of 10ppm of p-nitrophenol and at 1ppm of HgCl2.

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